Decellularized testicular matrix (DTM) enables researchers to focus on the specific composition of the testicular extracellular matrix (ECM) and elucidate its role in spermatogenesis. Furthermore, it provides the natural architectural arrangement that could guide the reorganization of dissociated testicular cells in vitro. This is a key consideration as the presence of an authentic nutritive and endocrine support has been proven to be essential for in vitro spermatogenesis, at least in the mouse (Oliver and Stukenborg in Andrology 8:825–834, 2020; Richer et al. in Andrology 12741, 2019). Hence, scaffolds of DTM could be harnessed for the development of a human in vitro spermatogenesis culture system, which is a missing link in male fertility preservation and could be a possible treatment for nonobstructive azoospermia (Gassei and Orwig in Steril 105:256–266, 2016).